Division of Reproductive Science in Medicine, Department of Obstetrics and Gynecology, Robert H. Lurie Comprehensive Cancer Center, Northwestern University Feinberg School of Medicine, Chicago, Illinois.
Department of Surgery, Northwestern University Feinberg School of Medicine, Chicago, Illinois.
Endocrinology. 2018 Oct 1;159(10):3581-3595. doi: 10.1210/en.2018-00559.
Progesterone is a steroid hormone that plays an important role in the breast. Progesterone exerts its action through binding to progesterone receptor (PR), a transcription factor. Deregulation of the progesterone signaling pathway is implicated in the formation, development, and progression of breast cancer. Next-generation selective progesterone receptor modulators (SPRMs) have potent antiprogestin activity and are selective for PR, reducing the off-target effects on other nuclear receptors. To date, there is limited information on how the newer generation of SPRMs, specifically telapristone acetate (TPA), affect PR function at the molecular level. In this study, T47D breast cancer cells were used to investigate the molecular mechanism by which TPA antagonizes PR action. Global profiling of the PR cistrome and interactome was done with chromatin immunoprecipitation sequencing (ChIP-seq) and rapid immunoprecipitation mass spectrometry. Validation studies were done on key genes and interactions. Our results demonstrate that treatment with the progestin (R5020) alone resulted in robust PR recruitment to the chromatin, and addition of TPA reduced PR recruitment globally. TPA significantly changed coregulator recruitment to PR compared with R5020. Upon conservative analysis, three proteins (TRPS1, LASP1, and AP1G1) were identified in the R5020+TPA-treated group. Silencing TRPS1 with small interfering RNA increased PR occupancy to the known PR regulatory regions and attenuated the inhibition of gene expression after TPA treatment. TRPS1 silencing alleviated the inhibition of proliferation by TPA. In conclusion, TPA decreases PR occupancy on chromatin and recruits coregulators such as TRPS1 to the PR complex, thereby regulating PR target gene expression and associated cellular responses.
孕激素是一种在乳腺中发挥重要作用的甾体激素。孕激素通过与孕激素受体(PR)结合发挥作用,PR 是一种转录因子。孕激素信号通路的失调与乳腺癌的形成、发展和进展有关。新一代选择性孕激素受体调节剂(SPRMs)具有强大的抗孕激素活性,对 PR 具有选择性,减少了对其他核受体的脱靶效应。迄今为止,关于新一代 SPRMs(特别是醋酸他普瑞林(TPA))如何在分子水平上影响 PR 功能的信息有限。在这项研究中,使用 T47D 乳腺癌细胞来研究 TPA 拮抗 PR 作用的分子机制。通过染色质免疫沉淀测序(ChIP-seq)和快速免疫沉淀质谱对 PR 顺式作用元件组和互作组进行了全局分析。对关键基因和相互作用进行了验证研究。我们的结果表明,单独用孕激素(R5020)处理会导致 PR 大量募集到染色质上,而添加 TPA 会导致 PR 募集总体减少。与 R5020 相比,TPA 显著改变了 PR 的共激活因子募集。经过保守分析,在 R5020+TPA 处理组中鉴定出三种蛋白质(TRPS1、LASP1 和 AP1G1)。用小干扰 RNA 沉默 TRPS1 可增加 PR 对已知 PR 调控区域的占有率,并减弱 TPA 处理后基因表达的抑制。TRPS1 沉默减轻了 TPA 对增殖的抑制作用。总之,TPA 降低了 PR 在染色质上的占有率,并招募了共激活因子(如 TRPS1)到 PR 复合物中,从而调节 PR 靶基因的表达和相关的细胞反应。